bioluminous insects and its features

2. Gande sravan kumar
Roll No:M-1063/17

3. Contents
 Introduction
 History of Bioluminescence
 Significance of Bioluminescence
 Survey of Luminous insects
 Physiology of light emission in
Lampyridae
 Chemistry of Bioluminescence in
Lampyridae
 Applications of Bioluminescence
 Current projects
 Case studies
 Conclusion

4. Incandescence – light by heating
c
Luminescence - light other than by heating
Bioluminescence - light by a living
organism.
Introduction

5. Cont…
Cold light emission
Hybrid word,
Greek → Bios = Living
Latin → Lumen = Light
Also known as ‘Chemiluminescence’ .

6. No self -luminous in
flowering plants,
birds, reptiles and
mammals.
Non marine-less
wide in
distribution but
larger variety
colors
Widely
distributed in
marine
organisms
but mostly
blue and
green lights
Seen in
Jelly fish Marine octopus
Panellus stipticus)

7.  Unique – color, wavelength, timing & interval
 Throughout the biosphere, but only at levels
below the mammals & plants
 17 phyla & 700 genera
 Terrestrial organisms (Annelids, Arthropods,
Molluscs), Fungi, and microorganisms

8. Collembola Hemiptera Coleoptera
Diptera
Insects have received much attention

9. History of
Bioluminescence

10. 1000 BC-Chinese book of odes
Aristotle (384-322 BC) –cold light
Japan the light of firefly-symbol of the souls
of the recently deceased(the Hitodama).

11.  Greeks and Romans - characteristics of luminous organisms
 1555 – first book devoted to Bioluminecsence – Conrad Gesner
 1667 – Robert Boyle – air requirement
 1873 - Raphael Dubois – extracted & coined the terms "luciferine"
and "luciferase".
 1959 – Harvey – Luciferin – luciferase system
Cont…

12. Mating signal
Predation
Defense
Illumination
Significance
(Gajendra Babu & Kannan., 2002 )

13.  Eg :- Fireflies
 Same species to aggregate
 Two communication systems :
1. wingless & sedentary females(Lampyris spp)
2. Photinus spp & Photuris spp
 Male fireflies seeking mates
 Female : male – 1 : 50
 Flash pattern vary with species & sex
 Flashes begin / end sharply
Mating signal

14. Photuris pyralis :-
500 ms duration flash at 6 s interval
Climb steeply & hovers for 2 s
Female flash 1.5 to 2.5 s after male flash
Male fly towards female
Flash again 3 s later
( Llod., 1971 , Sivinsky, 1981)

15.  New Zealand glow worm fly ( Arachnocampa luminosa
(Diptera: Keroplatidae)
 Female fly – egg – ceiling of dark caves in Waitomo
 Prey Eg :- Chironomid midges
 Luminous caves ( tourist attraction spots in New Zealand )
 Show route
(Gajendra Babu & Kannan., 2002 )
Predation

16. Photuris spp :
 Female imitate pattern of females of other species
(mostly Photinus spp.)
 Lure males of those species
 Feed on them ( femme fatale)
( Vencl et al., 1994 )

17.  Eg :- Rail road worms
(Order: Coleoptera)
 Lateral light organs
 Live in high densities
 Sudden flashes
 Repel potential predators
 Intimate mated females – lay eggs – over crowding
& competition for food source
( Gajendra Babu & Kannan., 2002 )
Defence

18.  Eg :- Rail road worms
 Head light organs
 Red light ( long wavelength )
 Not visible to prey
 Search for millipedes
(Viviani & Bechara., 1997 )
Illumination

19. Survey
 COLLEMBOLA (spring tails)
 DICTYOPTERA ( cockroaches )
 HEMIPTERA
Fulgoridae – lantern flies
 DIPTERA ( Glow worm flies )
Bolitophilidae
Platyuridae
 COLEOPTERA ( Beetles )
Elateridae - click beetles
Lampyridae - fireflies
Phengodidae - railroadworms

20.  c
Adult
Eg :- Onychiurus
armatus
Soil
Greenish flash throughout
the body
Fat body
Stimulation is required
Significance unknown
(rare)
Collembola
Eg: Giant cockroach
(Blaberidae)
Luchhormetica luckae
 Recently found in the
Amazon forest.
 It mimics click beetle
( Peter Vršanský et al 2012)
DICTYOPTERA
Fulgora laternaria (F :- Fulgoridae)
Adults
Both sexes should be together
Mating signal
No work on physiology &
biochemistry
Hemiptera : lantern flies

21. Diptera
Glow worm flies - F:- Bolitophilidae
Eg :- Arachnocampa luminosa or
Bolitophila luminosa
Excretory organ – photogenic function
Larvae – blue to green
Female – weakly luminescent
Male – loose after emergence from pupae
Eg :- Orfelia fultoni F : Mycetophilidae
Larva – Blue color
Webs – along stream banks in the Appalachian mountains
Lure prey
Light organ – anterior part & in tail

22. Eg:- Keroplatus sesoides
 Real fungus gnat
 Eat fungal spores (Oospores)
 No special light organ
 Hypodermal fat body – stimulated
 Whole body of larva & pupa is
luminous

23. COLEOPTERA ( Beetles ) :
Most of the bioluminescent species
Super family :- Elateroidea
F :- Lampyridae ( fireflies & glow worms )
Larvae & adult – luminous
Yellow – green flashes – ventral lanterns
Species – specific signal ( adult )
Aposematism ( warning signal ) – larvae – abdominal
lanterns

24.  Green glow – prothoracic lanterns – disturbed –
defense
 Yellow – orange glow – abdominal lanterns – flying
 Luminous termite mounds – Brazilian savanna spp
(attract flying prey )

25. F :- Phengodidae ( Railroadworms )
 Male – not luminous
 Larvae & female – exuberant
Phrixothrix spp
 Lateral lanterns – yellow green
 Head lanterns – red
 Only terrestrial organism producing red light

26. Variation in Colour of Light
 Varies with species, environmental factors or structure of luciferase
Collembola – greenish flash throughout the body
Fulgora (Hemiptera) _ white
Most insects - yellow-green as in Photinus and Lampyris (Coleoptera).
In larval and adult female railroad worms, thorax and abdomen – green to
orange. head - red light
Arachnocampa - blue-green

28. Physiology of light emission in Lampyridae
 Light organ – photophore / lantern
May occur in both sexes Eg :- Fire flies
Photuris spp ( ventral side )
Males -2pairs
Female -1pair

29.  Rail road worms :
(larvae & female )
11 pairs-dorso lateral
side of thorax and
abdomen
1 pair on head
 Fulgora - 1 light organ on head

30. Simplest case :-
Eg :- Phengodid larvae
 Gaint oenocyte like cells
 Lack neural control ( slow responding light system )
Specialized case :-
Eg :- Photinus spp & Photuris spp
 Rousettes of thousands of photocytes interpenetrated by
tracheaeoles

31.  Photocytes – lie beneath
epidermis
 Cuticle – transparent (
permit light )
 Trachea and nerves
perpendicular to cuticle
 Tracheal end cell –
sphincter (neuralcontrol)
 Tracheolar cell
 Neuron – spatulate
terminal processes

32. 32

33.  3 substances are involved
(Raphael Dubois., 1885 )
 Luciferin – heat resistant substrate & source of
light
 Luciferase – enzyme – trigger
 Molecular oxygen – fuel
 ATP, Mg+2 ions – catalysts
 Chemical reaction
occurs in cytoplasm
Chemistry

34.  Luciferin – low molecular weight compound – an
aldehyde, a polypeptide complex or a protein
Sequence of events involved in light production in insects

35. Importance
Besides their beauty and scientific interest - very important
to the society
luciferases and luciferins - sensitive tools - biotechnology
and biomedicine.
luciferase genes - bioluminescent markers - tuberculosis and
HIV
Biosensors - pollutants and environmental disruptors

36. Applications
Space Research
Pest management
Fluorescent marker gene
Medical research

37. Space Research
 Luciferin & luciferase systems – spacecrafts – Mars
 Existence of life forms
Special electronic device
Pick up soil
Mix with water, oxygen, luciferin & luciferase
Glow
 ATP ( 5th requirement )
 one quadrilionth of a gm - sensitive

38. Pest management
 Organisms distribution pattern
 USA, 2001 – GFP – jelly fish ( Aequora victoria )
 Modified genetic material of Pink boll worm
 Transgenic PBW strain ( green flourescent larvae )
 Objective – 2 fold
1. Distribution pest
2. Temperature sensitive lethal gene
( management )

39. Fluorescent marker gene
 Marker gene – short sequence of DNA
 Acts as a label
 Inserted along with a gene of interest into
cells
 Fluorescent marker genes – transformed
cells glow under light
 Help scientists – select transformed cells

40. Medical research
 Luciferase is injected into cells which glow
 Monitor progress in treatment as cells glowing
begin to disappear
 Muscular dystrophy, heart disease, urology,
antibiotic testing and waste water treatment
 Blood banks have used the luciferase enzyme to
test the quality of red blood cells within their
stock.

41. How Luciferin Help Us Understand Cancer
Normal cells
 Respond to growth
factors ( signals )
 Food & nutrients –
instructions
 Excess growth & cell
division is avoided
Cancer cells
 Ignore signals
 Uncontrolled uptake
of food & nutrients
due to mutation in
genes
 Tumor formation

42.  1924, Otto Warburg
 Glucose – cancer cells >
normal cells
 Convert to Lactate
 Plenty of oxygen – aerobic
glycolysis
 Altered metabolism –
Warburg effect
Tumor cells produce lactate
even when oxygen is present

43. The firefly "Christmas Tree" as observed in regions of S.E Asia, from Malaysia to
Papua-New Guinea. It is due to the synchronous flashing of fireflies.

44. Image of bioluminescent red tide event of 2005 at a beach in Carlsbad California
showing brilliantly glowing crashing waves containing billions of
Lingulodinium polyedrum dinoflagellates. The blue light is a result of a luciferase enzyme.

45. Current projects
 Structure / Function relationship of insect
luciferase
 Molecular origin and evolution of insect luciferase
 Biodiversity and photoecology of terrestrial
bioluminescence
Prof. Dr. Vadim Viviani,
Departamento de Biologia, Brazil.

47. Construction, characterization and exemplificative
application of bioluminescent
Bifidobacterium longum (Guglielmetti s. et al,2008).
Department of Food Science and Microbiology,
University of Milan, Italy
Institute of Environmental Engineering and
Biotechnology,Tampere University of Technology, Finland

48. AIM : To construct a bifidobacterial luminescent biosensor
that could be used for a quick analysis of the metabolic state
of cells under different conditions.
RESULTS : Detected a minimum of 4000 cells, which indicates
that the insect luciferase expression in Bifidobacterium longum is
extremely good, and a measurement requires only a few minutes
of incubation

49. ATP-Bioluminescence as a method to evaluated
microbiological quality of UHT milk (A.F. Cunha et
al,2014).
LOCATION :
Brazil United States – St. Paul, MN, United States.

50. To compare the results of culturemethods with the
results of ATP-Bioluminescence technique of 102 UHT
whole milk samples incubated at 48, 72, and 168 hours.
RESULT :
Analyzed for the presence of mesophilic and psychrotrophic
aerobic microorganisms using Plate Count Agar (PCA), Brain-Heart
Infusion (BHI)media and PetrifilmTM Aerobic Count (AC) plate.
For the dairy industry, the ATP-Bioluminescence techniquemay
become an important tool that assists the official methods to
quickly monitor the microbiologicalquality of UHT milk though this
will likely require a threshold below 150 RLU.

51. Conclusion :-
 Bioluminescence - light by a living organism (
cold light )
 2000 lumiscent species in insects
 Functional significance ( mating signal, defence,
predation ) – biological control of pests
 Luciferase – sensitive tool in biotechnology,
biomedicine
 Biosensors – pollutants & environmental disruptors